Within the current context of declining biodiversity, the botanical gardens play an essential role in its conservation. Dianthus callizonus, D. glacialis ssp. gelidus and D. spiculifolius are the species that we seek to preserve in "Alexandru Borza" Botanical Garden of Cluj-Napoca (Romania). Several replicates were collected for each taxon from different populations in order to avoid the genetic uniformity. The material collected from the natural sites, was planted on a rockery, specially designed for this collection in the Botanical Garden. At the time of planting, each individual was sampled for setting up an in vitro collection and further biochemical and molecular analyses. In case of ex situ outdoor conservation of the three Dianthus species, 80.6% of the individuals collected in the field survived during the first year but the percentage decreased drastically after four years. In the case of in situ collected individuals, as well as in the case of in vitro individuals, D. spiculifolius had the best ability to acclimatize in the Botanical Garden, and D. callizonus presented the lowest number of surviving individuals. The ex vitro acclimatization of the plantlets had 80% efficiency at 10ÂºC, using three different substrates: soil and pearl stone mix 1/1, soil and sand mix 1/1 and pearl stone. All the three species are preserved in vitro, whereas the plantlets are acclimatized outdoors. Ex situ conservation of these species will have a positive impact on the biodiversity conservation.